Cense Biosciences was formed specifically to create a minimally invasive, self-regulating delivery technology for insulin, and to extend this technology to other active drugs. The core technology of CenseI, AVT, or Agglomerated Vesicle TechnologyII, developed by the Chief Scientist of Cense, is a chemically cross linked agglomerate of drug encapsulating nanoparticles (such as liposomes) that modulates release by cleavage of the cross links. Insulin is contained within the nanoparticles, while the cross links are capable of cleavage as required. Thus, hyperglycemic events trigger insulin release from the particles, causing an automatic return to normoglycemia. In addition the obvious advantages of autoregulated delivery, the key additional advantage of this technology are the active drug is not chemically modified, thus avoiding the issues surrounding an NCE. The major shortcoming of the current AVT however, is that the glucose-sensitive linkage is based on competitive binding of concanavalin-A (Con-A) to a sugar. Con-A is a lectin, with high glucose affinity, but is highly inflammatory. In this Phase 1 proposal therefore, we propose to identify a suitable glucose binding replacement for Con-A and then test the performance of AVT particles made using this new molecule. i A. Annapragada, R. Bhavane, Agglomerated particles for aerosol drug delivery, US patent application 20030190284, October 9, 2003. ii E. Karathanasis, R. Bhavane, A. V. Annapragada, Triggered release of inhaled insulin from the agglomerated vesicles: Pharmacodynamic studies in rats. Accepted for publication in Jour. Of Controlled release April 2006. PUBLIC HEALTH RELEVANCE: Cense Biosciences was formed specifically to create a minimally invasive, self-regulating delivery technology for insulin, and to extend this technology to other active drugs. The core technology of CenseI, AVT, or Agglomerated Vesicle TechnologyII, developed by the Chief Scientist of Cense, is a chemically cross linked agglomerate of drug encapsulating nanoparticles (such as liposomes) that modulates release by cleavage of the cross links. Insulin is contained within the nanoparticles, while the cross links are capable of cleavage as required. Thus, hyperglycemic events trigger insulin release from the particles, causing an automatic return to normoglycemia. In addition the obvious advantages of autoregulated delivery, the key additional advantage of this technology are the active drug is not chemically modified, thus avoiding the issues surrounding an NCE. The major shortcoming of the current AVT however, is that the glucose-sensitive linkage is based on competitive binding of concanavalin-A (Con-A) to a sugar. Con-A is a lectin, with high glucose affinity, but is highly inflammatory. In this Phase 1 proposal therefore, we propose to identify a suitable glucose binding replacement for Con-A and then test the performance of AVT particles made using this new molecule. We will begin by screening a family of boronates, known glucose-binding molecules, some of which are already used in vivo. Within this Phase 1 project, we will identify new glucose-binding molecules for AVT, using high-content and high-throughput assay methods. Glucose binding will be measured by a competitive sugar binding assay. Inflammation will be measured by an imaging assay for NF: B nuclear translocation. After identification of lead candidates, we will produce AVT particles with them, and test them in vitro, and compare their behavior with the gold-standard ConA particles. In vitro cytotoxicity will also be assessed. The key milestone for Phase 1 will be the identification of 1-10 boronate compounds that have lower inflammatory characteristics than ConA, and equivalent in vitro AVT performance. Upon moving into Phase 2, AVT particles will be tested in normal and diabetic rat models. Pharmacodynamic, pharmacokinetic and inflammation properties of the preparations will be tested, and lead candidates selected for further development. The novelty of this project lies in 2 key areas: (1) the development of a truly "smart" insulin delivery system (2)the use of drug screening methods (HTS, HCS) for the screening of recipients. Both of these are landmarks in the pharmaceutical industry. I A. Annapragada, R. Bhavane, Agglomerated particles for aerosol drug delivery, US patent application 20030190284, October 9, 2003. ii E. Karathanasis, R. Bhavane, A. V. Annapragada, Triggered release of inhaled insulin from the agglomerated vesicles: Pharmacodynamic studies in rats. Accepted for publication in Jour. Of Controlled release April 2006.